Mechanism of bone marrow mesenchymal stem cell exosomes regulating NLRP3 inflammatory bodies on alveolar bone defect in diabetes rats
Objective To explore the molecular mechanism of bone marrow mesenchymal stem cell exosomes on alveolar bone defect healing in diabetic rats.Methods A total of 50 SD rats were selected to be included in this study,and 10 rats were randomly selected as the control group.The remaining rats were used to construct the diabetes plus bilateral maxillary alveolar bone defect model.The model group,BMMSC-Exos group,MCC950 group(NLRP3 inhibitor),BMMSC-Exos and MCC950 group were set up.The BMMSC-Exos and MCC950 group of SD rats were injected with BMMSC-Exos and MCC950 reagents through the tail vein,while the model group was injected with an equal amount of saline;After 28 days of model construction,the maxilla tissue of rats was taken.TEM was used to detect the morphology of BMMSC-Exos,and Western blot was used to detect the expression of BMMSC-Exos marker proteins CD9,CD63,and CD81.Fasting blood glucose levels,HE staining analysis of alveolar bone inflammatory infiltration and Lance-Sandhu score in each group of rats were analyzed;The NLRP3 inflammasome signaling pathway key proteins(NLRP3,caspase-1,IL-1β)in alveolar bone tissue of rats in each group,and the expression of key proteins in bone metabolism(OPG,ALP,Collal)were analyzed by Western blot.Results The diameter of BMMSC-Exos is around 100 nm,presenting a double concave disc state.Compared with pure BMMSCs,the membrane structure is intact,and the expression of BMMSC-Exos marker proteins CD9,CD63,and CD81 was significantly increased(P<0.05).The model of diabetic rats was successfully established.BMMSC-Exos intervention and NLRP3 inhibitor(MCC950)reduced the blood glucose level of the rats(P<0.05);and combination of the two treatments further reduced the blood sugar level of the model rats(P<0.05).Compared with control group,the bone regeneration Lane-Sandhu score,OPG,ALP,Collal expression were down-regulated,while the NLRP3,caspase-1,IL-1β expression levels were up-regulated,in the diabetic alveolar bone defect model(P<0.05).BMMSC-Exos intervention greatly up-regulated the Lane-Sandhu score and the expression of OPG,ALP,and Collal proteins(P<0.05),and down-regulated the NLRP3,caspase-1,and IL-1β expression(P<0.05).Compared with the model group,BMMSC-Exos and MCC950 intervention increased the Lane-Sandhu score of bone regeneration and the expression of key bone metabolism proteins(OPG,ALP,Collal)in rats(P<0.05).Combination of the two treatments further up-regulated the Lane-Sandhu score and expression of key bone metabolism proteins in rat bone regeneration(P<0.05).Conclusion The exosomes of bone marrow mesenchymal stem cells can benefit the bone metabolism of diabetic rats with alveolar bone defect by inhibiting the activity of NLRP3 inflammasome,reducing the inflammatory reaction and promoting the healing of bone defect.
extracellular vesiclesbone marrow mesenchymal stem celldiabetesalveolar bone defectNLRP3 inflammasomeheal
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